Methods of using therapeutic phospholipid derivatives

ABSTRACT

A method of treating protozoal and fungal diseases is described, in which an effective amount of a compound of formula I is administered to a host having a protozoal or fungal disease. Also described are methods of treating bone marrow damage composed of administering, to a host having bone marrow damage due to treatment with cytostatic agents or other myelotoxic active ingredients, an effective amount of a compound of formula I

This is a division of application Ser. No. 08/905,333, filed Aug. 4,1997 which is allowed U.S. Pat. No. 6,172,050 which is a cont. of Ser.No. 08/487,624 filed Jun. 7, 1995 abandoned, which is a cont. of Ser.No. 08/086,850, filed Jul. 7, 1993 abandoned.

BACKGROUND OF THE INVENTION

The present invention relates to Published European Patent Application108 565 relates to compounds of the General Formula:

and their pharmaceutically acceptable salts, in which R¹ is an aliphatichydrocarbon radical having 8-30 carbon atoms and the radicals R², R³ andR⁴ are identical or different and are hydrogen or lower alkyl radicals,or in which the group NR²R³R⁴ is a cyclic ammonium group, and n has thevalue 0 or 1. Antitumor and antifungal activity are indicated for thesecompounds.

SUMMARY OF THE INVENTION

The present invention relates to alkyl or alkene phosphates in which thecholine radical is part of a heterocyclic ring, to a process for thepreparation of the class of compounds, to pharmaceutical compositionscontaining the compounds as active ingredients and to processes for thepreparation of said drugs.

More specifically, the present invention provides compounds of theGeneral Formula I:

in which

R is a linear or branched alkyl radical having 10 to 24 carbon atoms,which can also contain one to three double or triple bonds, R¹ and R²independently of one another are hydrogen or in each case a linear,branched or cyclic saturated or unsaturated alkyl radical having 1 to 6carbon atoms, which can also contain a Cl, OH or NH₂ group, it alsobeing possible for two of these radicals to be bonded together to form aring,

A is a single bond or one of the groups of the formulae

—CH₂—CH₂—CH₂—O—  (II),

—CH₂—CH₂—O—  (III),

 the groups (II) to (VI) being orientated in such a way that the oxygenatom is bonded to the phosphorus atom of compound (I), X is an oxygen orsulphur atom or NH when A is a single bond, or an oxygen or sulphur atomwhen A is one of the groups (II) to (VI),

y is equal to 0 or a natural number between 1 and 3, and

m and n independently of one another are 0 or natural numbers, with theproviso that m+n=2 to 8.

The present invention also provides a pharmaceutical compositioncomprising, as the active ingredient, at least one compound according toGeneral Formula (I) or a pharmaceutically acceptable salt thereof and apharmaceutically acceptable carrier therefor. The pharmaceuticalcomposition may also include pharmaceutically acceptable excipients,adjuncts, fillers and diluents. The amount of active ingredient in thepharmaceutical dosage unit the pharmaceutical composition is preferablybetween 50 mg and 250 mg. Preferred compounds for the pharmaceuticalcomposition are selected from the group consisting of octadecyl1,1-dimethylpiperidinio-4-yl phosphate, octadecyl1,1-dimethylperhydroazepinio-4-yl phosphate, octaecyl1,1-dimethylperhydroazepinio-4-yl phosphate, erucyl1,1-dimethylpiperidinio-4-yl phosphate and erucyl1,1-dimethylperhydroazepinio-4-yl phosphate.

The present invention also provides methods of treating a tumor,autoimmune disease or skin disease or skin disease, and of combatingprotozoal and fungal diseases, which comprises administering to a hostin need o such treatment an effective amount of a compound of GeneralFormula (I). Such methods are particularly useful for treatingleishmaniasis, multiple sclerosis, and psoriasis.

In addition, the invention provides a method of treating bone marrowdamage due to treatment with cytostatic agents and other myeloxtoxicactive ingredients which comprises administering, to a host having bonemarrow damage due to treatment with cytostatic agents or othermyelotoxic active ingredients, an effective amount of a compound ofGeneral Formula (I).

The invention also provides a method of treating a viral disease whichcomprises administering to a host having such a disease an effectiveamount of a compound of General Formula (I). This method should beparticularly useful in treating AIDS.

Surprisingly, the compounds according to the invention have betterantitumor activity than the open-chain derivatives described in EP-A 108565. The invention further relates to processes for the preparation andprocesses for the purification of the novel compounds.

More specifically, the present invention relates to a procedure for thepreparation of compounds of general formula I—further referred to asprocess A—in which a compound of the general formula VII.

R—X—A—H  (VII)

in which R, X and A are as defined above, is reacted with phosphorusoxytrichloride in the presence of a suitable auxiliary base, with orwithout a solvent, and then reacted with a compound of the generalformula:

in which R₁, R², y, m and n are as defined above and Y⁻ is halide,mesylate or tosylate, to give compounds of the general formula I, oroptionally compounds of the General Formula IX:

in which R¹, y, m and n as defined above can be used instead ofcompounds of the general formula VIII during the process mentionedabove. Process B consists in the subsequent alkylation of compounds ofgeneral formula I obtained by process A, in which R¹ and/or R² arehydrogen, using alkylating agents R²—Y in which R² is as defined aboveand Y is chlorine, bromine, iodine, tosyl or mesyl, in a manner knownper se.

The present invention also provides a process for the purification ofthe compounds of General Formula I in which a solution of the compoundsof General Formula I, which have been prepared by means of knownprocesses or by a process as described above, in an organic solvent istreated with a mixed bed ion exchanger or successively or simultaneouslywith an acid or basic ion exchanger.

The first step of process A consists in reacting phosphorusoxytrichloride with a compound of Formula VII in halogenatedhydrocarbons, saturated cyclic ethers, acyclic ethers, saturatedhydrocarbons having 5 to 10 carbon atoms or liquid aromatic hydrocarbonswhich can also be substituted by halogen (especially chlorine), or inmixtures of the above-mentioned solvents, or without a solvent,optionally in the presence of a basic substance conventionally used forthis purpose.

Examples of possible halogenated hydrocarbons are hydrocarbons having 1to 6 carbon atoms, one or more or all of the hydrogen atoms beingreplaced with chlorine atoms. Methylene chloride, chloroform, ethylenechloride, chlorobenzene and dichlorobenzene, for example, can be used.In the case of halogen-substituted aromatic hydrocarbons, these arepreferably substituted by one or two halogen atoms.

Examples of saturated cyclic ethers which can be used are ethers with aring size of 5-6 which consist of carbon atoms and one or 2 oxygenatoms, examples of such ethers being tetrahydrofuran and dioxane.

The acyclic ethers consist of 2 to 8 carbon atoms and are liquid,possible examples being diethyl ether, diisobutyl ether, methyltert-butyl ether and diisopropyl ether.

Possible saturated hydrocarbons are unbranched and branched hydrocarbonswhich consist of 5 to 10 carbon atoms and are liquid, possible examplesbeing pentane, hexane, heptane and cyclohexane.

Examples of possible aromatic hydrocarbons are benzene andalkyl-substituted benzenes, the alkyl substituents consisting of 1 to 5carbon atoms.

Possible basic substances both for the reaction of the phosphorusoxychloride with the long-chain alcohol and for the subsequentconversion to the phosphoric acid diester are amines, for examplealiphatic amines of the formula NR₁R₂R₃, R₁, R₂ and R₃ being identicalor different and being hydrogen or C₁-C₆-alkyl, or else aromatic aminessuch as pyridine, picoline and quinoline. The basic substance requiredfor the conversion to the phosphoric acid diester can be addedsimultaneously with or before the amino alcohol or ammonium alcoholsalt.

A solvent is necessary in every case for the second reaction, i.e., ifthe first reaction step is carried out without a particular solvent, onemust now be added. The molar ratio of phosphorus oxychloride to thelong-chain alcohol is for example between 1.5:1 and 0.8:1.

The amino alcohol or the ammonium alcohol salt is for example used inexcess, based on the long-chain alcohol (about 1.1-1.5 molar excess).

If the reaction of the phosphorus oxychloride with the long-chainalcohol is carried out in the presence of a basic substance, the amountof the basic substance is for example 1 to 3 mol, based on 1 mol ofPOCl₃. The amount of basic substance used for the subsequent conversionto the phosphoric acid diester is for example 1 to 5 mol, based on 1mol.

The temperature of the reaction of phosphorus oxychloride with thelong-chain alcohol is between −30° C. and +30° C., preferably between−15° C. and +5° C. and especially between −10° C. and −5° C.

The duration of this reaction is for example 0.5-5 hours, preferably 1-3hours and especially 1.5-2 hours. If it is carried out in the presenceof a basic substance, the reaction generally proceeds rapidly (about 30minutes).

The amino alcohol or the ammonium alcohol salt is then added in portionsor all at once. Possible ammonium alcohol salts are those with mineralacids (for example sulphuric acid, hydrochloric acid) and also thosewith organic acids, for example acetic acid, paratoluene-sulphonic acidand the like. This reaction step takes place in an inert solvent.Possible solvents for this step are the same ones as those used for thereaction of the phosphorus oxychloride with the long-chain alcohol, inthe case where this reaction is carried out in a solvent..

The basic substance is then added dropwise, either dissolved in one ofthe indicated solvents or without a solvent. The following arepreferably used as solvents for the basic substance: halogenatedhydrocarbons, saturated cyclic ethers, acyclic ethers, saturatedhydrocarbons having 5 to 10 carbon atoms, liquid aromatic hydrocarbonsor mixtures of the above-mentioned solvents.

These are the same solvents as those which can be used for the reactionof the phosphorus oxychloride with the long-chain alcohol.

The addition of the basic substance raises the temperature. Care istaken to ensure that the temperature is kept in a range of between 0° C.and 40° C., preferably between 10° C. and 30° C. and especially between15° C. and 20° C.

The reaction mixture is then stirred at 5° C. to 30° C., preferably 15°C. to 25° C. (for example for 1 hour to 40 hours, preferably 3 hours to15 hours).

The reaction mixture is hydrolyzed by the addition of water, duringwhich the temperature should be kept at between 10° C. and 30° C.,preferably between 15° C. and 30° C. and especially between 15° C. and20° C.

The above-mentioned hydrolyzing liquids can also contain basicsubstances, such basic substances possibly being alkali metal andalkaline earth metal carbonates and bicarbonates.

To complete the hydrolysis, stirring is then continued for a further 0.5hour to 4 hours, preferably 1 to 3 hours and especially 1.5 to 2.5hours, at 10° C. to 30° C., preferably at 15° C. to 25° C. andespecially at 18° C. to 22° C.

The reaction solution is then washed with a mixture of water andalcohols (preferably saturated aliphatic alcohols having 1 to 4 carbonatoms) which can optionally also contain a basic substance. The mixingratio water:alcohol can be for example between 5 and 0.5, preferably 1-3(v/v).

Examples of possible basic substances for the washing liquid are alkalimetal and alkaline earth metal carbonates and bicarbonates, as well asammonia (for example aqueous ammonia). A 3% solution of sodium carbonatein water is particularly preferred.

The reaction solution can then optionally be washed with an acidsolution. The acid washing is advantageous for removing basic componentsof the reaction solution which have not yet reacted, especially whenmethylene chloride is used as the solvent.

The washing solution consists of a mixture of water and alcohols.Mixtures of saturated aliphatic alcohols having 1 to 4 carbon atoms arepreferred, it optionally being possible for an acid substance to bepresent as well. The mixing ratio water:alcohol can be for examplebetween 5 and 0.5, preferably 1-3 (v/v).

Examples of possible acid substances for the washing liquid are mineralacids and organic acids, for example hydrochloric acid, sulphuric acid,tartaric acid or citric acid. A 10% solution of hydrochloric acid inwater is particularly preferred.

This is followed by a further washing with a mixture of water andalcohols. Mixtures of saturated aliphatic alcohols having 1 to 4 carbonatoms are preferred, it optionally being possible for a basic substanceto be present as well. The mixing ratio water:alcohol can be for examplebetween 5 and 0.5, preferably 1-3.

The washed phases are then combined and dried in conventional manner,after which the solvent is removed (preferably under reduced pressure,for example at 5 to 100 mbar), optionally after the addition of 150-1000ml, preferably 300-700 ml and especially 450-550 ml of an aliphaticalcohol (based on 1 molar part by weight of dry product). Preferredalcohols are saturated aliphatic alcohols with a chain length of 1 to 5carbon atoms, particularly preferred alcohols being n-butanol andisopropanol. The purpose of this alcohol treatment is the completeremoval of residual water and the avoidance of foaming.

Further purification of the product can be effected for example bydissolving the crude product in hot ethanol, filtering off the residueand treating the filtrate with a mixed bed ion exchanger such as, forexample, Amberlite MB3 in ethanolic solution. Any commercially availableacid and basic ion exchangers can be used, simultaneously orsuccessively, instead of a mixed bed ion exchanger.

The solution is then recrystallized from ketones such as, for example,acetone or methyl ethyl ketone; digestion with the above solvents issufficient in some cases. It may be convenient to purify the products bycolumn chromatography or flash chromatography on silica gel usingmixtures of chloroform, methylene chloride, methanol and 25% ammoniasolution, for example, as the eluent.

Process variant B consists in the subsequent alkylation of productswhich are obtainable by process A using amino alcohols. Examples ofalkylating agents which can be used are methyl p-toluenesulphonate ordimethyl sulphate. Possible solvents are those which have been mentionedabove.

Alkali metal carbonates are examples of basic substances used. Thereaction is carried out at elevated temperature, for example at theboiling point of the solvents.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates Example 1 in the DMBA induced tumor-model, 4* per os,treatment day 0, 3, 7, 10.

FIG. 2 illustrates Example 1 in the DMBA induced tumor-model, 14* peros, treatment day 0-13.

FIG. 3 illustrates Example 1 in the DMBA induced tumor-model, ‘largetumors’, treatment day 0-27.

FIG. 4 illustrates Example 8 in the DMBA induced tumor-model, 4* per os,treatment day 0, 3, 7, 10.

FIG. 5 illustrates Example 13 in the DNBA induced tumor-model, 14* peros, treatment day 0-13.

FIG. 6 illustrates Example 13 in the DMBA induced tumor-model, ‘largetumors’, treatment day 0.

FIG. 7 illustrates Example 20 in the DMBA induced tumor-model, 4* peros, treatment day 0, 3, 7, 10.

FIG. 8 illustrates Example 21 in the DMBA induced tumor-model, 4* peros, treatment day 0, 3, 7, 10.

FIG. 9 illustrates Example 22 in the DMBA induced tumor-model, 4* peros, treatment day 0, 3, 7, 10.

FIG. 10 illustrates Example 1 in the KB tumor-model, 1* per os,treatment day 0.

FIG. 11 illustrates Example 8 in the KB tumor-model, 2* per os,treatment day 0, 7.

FIG. 12 illustrates Example 10 in the KB tumor-model, 2* per os,treatment day 0, 7.

FIG. 13 illustrates Example 17 in the KB tumor-model, 2* per os,treatment day 0, 7.

FIG. 14 illustrates Example 21 in the KB tumor-model, 2* per os,treatment day 0, 7.

FIG. 15 illustrates Example 22 in the KB tumor-model, 2* per os,treatment day 0, 7.

FIG. 16 illustrates Example 24 in the KB tumor-model, 2* per os,treatment day 0, 7.

FIG. 17 illustrates Example 25 in the KB tumor-model, 2* per os,treatment day 0, 7.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

The following examples illustrate the invention.

EXAMPLES Example 1

Name (IUPAC Nomenclature)

4-(((Octadecyloxy)hydroxyphosphenyl)oxy)-1,1-dimethylpiperidiniumhydroxide internal salt

Abbreviated Name

Octadecyl 1,1-dimethylpiperidinio-4-yl phosphate

C₂₅H₅₂NO₄P (461.66).1/2H₂O

Preparation Variant A

10.3 ml (0.11 mol) of phosphorus oxychloride are placed in 100 ml ofchloroform and cooled to 5-10° C. A solution of 27.0 g (0.10 mol) of1-octadecanol in 100 ml of chloroform and 35 ml of pyridine is addeddropwise over 30 min, with stirring. After subsequent stirring for 30min at 5-10° C., 39.1 g (0.13 mol) of 4-hydroxy-1,1-dimethylpiperidiniumtosylate are added in a single portion. After the addition of 40 ml ofpyridine and 30 ml of DMF, the mixture is stirred for 24 hours at roomtemperature. It is then hydrolyzed with 15 ml of water and subsequentlystirred for 30 min and the organic phase is washed with 200 ml each ofwater/methanol (1:1), 3% Na₂CO₃/methanol (1:1) and finallywater/methanol (1:1). The organic phase is concentrated, the residue isdissolved in 300 ml of hot ethanol and the solution is filtered aftercooling. The filtrate is stirred with 80 g of Amberlite MB3 ionexchanger, the mixture is filtered and the filtrate is concentrated. Theresidue is recrystallized from 300 ml of methyl ethyl ketone, filteredoff with suction and dried under vacuum over P₂O₅.

Yield: 4.71 g (10%)

Elemental analysis:

C H N calc.: 65.26% 11.63% 2.62% found: 64.38% 11.61% 2.73% 65.04%11.80% 2.78%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.17

(1-butanol/glacial acetic acid/water 40:10:10)

Rf=0.12

Melting point: 270-271° C. (decomposition)

Preparation Variant B

20.1 ml (0.22 mol) of phosphorus oxychloride are placed in 100 ml ofmethylene chloride and cooled to 5-10° C. and a solution of 54.1 g (0.20mol) of octadecanol in 400 ml of methylene chloride and 70.5 ml ofpyridine is added over 30 min, with stirring. After subsequent stirringfor one hour, 29.9 g (0.26 mol) of 4-hydroxy-1-methylpiperidine in 80 mlof pyridine are added dropwise. After stirring for 3 hours at 10° C.,the mixture is hydrolyzed with 30 ml of water while being cooled withice and is subsequently stirred for one hour. The organic phase iswashed with 200 ml each of water/methanol (1:1), 3 percent hydrochloricacid/methanol (1:1) and water/methanol (1:1). The organic phase is driedover Na₂SO₄ and concentrated until turbidity appears, and 1 liter ofmethyl ethyl ketone is added. The crystals are recrystallized from 1liter of methyl ethyl ketone, filtered off with suction and dried undervacuum over P₂O₅.

Yield:

54.1 g (60%) of octadecyl 1-methylpiperidinio-4-yl phosphate

98.1 g (0.22 mol) of octadecyl 1-methylpiperidinio-4-yl phosphate aresuspended in 500 ml of absolute ethanol and heated to boiling. Underreflux, a total of 71.8 g (0.39 mol) of methyl p-toluenesulphonate and26.5 g (0.19 mol) of potassium carbonate are added alternately in eightportions over 2 hours. When the addition is complete, the mixture isrefluxed for a further hour. After cooling, it is filtered, the filtrateis concentrated to half and 150 g of moist Amberlite MB3 ion exchangerare added to the solution. After stirring for two hours, the mixture isfiltered with suction over kieselguhr/activated charcoal and thefiltrate is concentrated and crystallized with acetone. The crystal cakeis recrystallized from methyl ethyl ketone and dried under vacuum overP₂O₅.

Yield: 46.1 g (46%) of octadecyl 1,1-dimethylpiperidinio-4-yl phosphate

Elemental analysis:

C H N calc.: 65.26% 11.63% 2.62% found: 65.18% 11.62% 2.68% 65.07%11.71% 2.70%

Melting point: 271-272° C. (decomposition)

Example 2 Hexadecyl piperidinio-4-yl phosphate

C₂₁H₄₄NO₄P (405.558),

7.1 ml (77 mmol) of phosphorus oxychloride are dissolved in 50 ml of drytetrahydrofuran and, after cooling to 5-10° C., a solution of 17 g (70mmol) of hexadecanol and 48 ml of triethylamine in 150 ml oftetrahydrofuran is added dropwise, with stirring. When the addition iscomplete, the mixture is subsequently stirred for 30 min in an ice bathand then left to warm up to room temperature. 10.1 g (100 mmol) of4-piperidinol are dissolved in 100 ml of tetrahydrofuran and mixed with17 ml of triethylamine and the mixture is added dropwise to the reactionsolution, with stirring, so that the temperature does not exceed 40° C.When the addition is complete, the mixture is refluxed for one hour.While still hot, the solution is separated from the triethylammoniumchloride by filtration and, after cooling, is poured into an ice/2 Mhydrochloric acid mixture, with stirring. The product obtained oncooling in a refrigerator is taken up in methylene chloride, dried overMgSO₄, concentrated and chromatographed on silica gel with methylenechloride/methanol/25% ammonia (70:30:5). The product fractions arecombined and concentrated. After recrystallization from methanol, theproduct is dried under vacuum over P₂O₅.

Yield: 10.0 g (35%)

Elemental analysis:

C H N calc.: 62.19% 10.94% 3.45% found: 65.15% 11.14% 3.54% 62.41%11.19% 3.34%

Thin-layer chromatogram

(chloroform/methanol/25% ammonia 70:20:10)

Rf=0.42

(1-butanol/glacial acetic acid/water 40:10:10)

Rf=0.33

Example 3 Hexadecyl 1,1-dimethylpiperidinio-4-yl phosphate

C₂₅H₅₂NO₄P (461.64).H₂O

5.7 g (14 mmol) of hexadecyl piperidinio-4-yl phosphate are dissolved in100 ml of methanol and mixed with 11.6 g (84 mmol) of potassiumcarbonate. 4.0 ml (42 mmol) of dimethyl sulphate are added dropwise over30 min, with thorough stirring. The mixture is subsequently stirred for4 hours at 40° C., cooled, filtered and concentrated. The residue isdigested with acetone and, after filtration with suction, is dissolvedin 100 ml of 96% ethanol. 15 g of Amberlite MB3 ion exchanger are addedand the mixture is stirred for 3 hours. After filtration, the filtrateis concentrated and recrystallized twice from methyl ethyl ketone. Thecrystals are dried under vacuum over P₂O₅.

Yield: 3.70 g (61%)

Elemental analysis:

C H N calc.: 61.17% 11.16% 3.10% found: 60.83% 11.14% 2.99% 60.92%11.26% 3.00%

Thin layer chromatogram:

(chloroform/methanol/25% ammonia 70:20:10)

Rf=0.28

(1-butanol/glacial acetic acid/water 40:10:10)

Rf=0.13

Melting point: 230° C. (decomposition)

Example 4 Erucyl 1,1-dimethylpiperidinio-4-yl phosphate

C₂₉H₅₈NO₄P (515.765).H₂O

10.3 ml (0.11 mol) of phosphorus oxychloride are placed in 50 ml ofchloroform, and a solution of 32.5 g (0.10 mol) of erucyl alcoholcis-13-docosenyl alcohol and 32 ml of pyridine in 100 ml of chloroformis added dropwise at 5-10° C. After subsequent stirring for half anhour, 39.1 g (0.13 mol) of 4-hydroxy-1,1-dimethylpiperidinium tosylateare added in a single portion. After the dropwise addition of 40 ml ofpyridine, the mixture is left to warm up to room temperature and stirredfor 3 hours. It is then hydrolyzed with 15 ml of water, subsequentlystirred for half an hour and washed with 100 ml each of water/methanol(1:1), 3% sodium carbonate solution/methanol (1:1), 3% citricacid/methanol (1:1) and water/methanol (1:1). The residue obtained afterconcentration of the organic phase is digested with acetone and thendissolved in 150 ml of 96% ethanol. This solution is stirred for 3 hourswith 20 g of Amberlite MB3 ion exchanger and filtered over kieselguhr togive a clear solution. This is concentrated and chromatographed onsilica gel with chloroform/methanol/25% ammonia 70:40:10. The productfractions are combined and concentrated to dryness under vacuum.

Yield: 4.4 g (9%)

Elemental analysis:

C H N calc.: 65.26% 11.63% 2.62% found: 64.38% 11.61% 2.73% 65.04%11.80% 2.78%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:20:10)

Rf=0.30

Example 5 Hexadecyl 1,1-dimethylpiperidinio-3-yl phosphate

C₂₃H₄₈NO₄P (433.616).H₂O

10.3 ml (0.11 mol) of phosphorus oxychloride are placed in 50 ml ofchloroform and cooled to 0-10° C. 24.2 g (0.10 mol) of n-hexadecanol aredissolved in 100 ml of chloroform, 32 ml of pyridine are added and themixture is added dropwise to the phosphorus oxychloride solution overone hour, with ice cooling. After subsequent stirring for half an hour,39.2 g (0.13 mol) of 3-hydroxy-1,1-dimethylpiperidinium tosylate areadded in a single portion and 40 ml of pyridine are added dropwise over15 min at room temperature. After stirring for 16 hours at roomtemperature, the mixture is hydrolyzed with 15 ml of water, stirred forhalf an hour and washed with 100 ml each of water/methanol (1:1), 3%sodium carbonate solution/methanol (1:1), 3% citric acid/methanol (1:1)and water/methanol (1:1).

The organic phase is dried over sodium sulphate and concentrated. Theresidue is dissolved in 150 ml of 96% ethanol, the solution is filteredand the filtrate is stirred with Amberlite MB3 ion exchanger. After theion exchanger has been filtered off, the filtrate is concentrated andthe residue is crystallized with acetone, filtered off with suction anddried under vacuum over P205.

Yield: 13.5 g (31%)

Elemental analysis:

C H N calc.: 61.17% 11.16% 3.10% found: 60.78% 11.41% 2.87% 60.85%11.31% 2.86%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.37

Example 6 Octadecyl 1,1-dimethylpiperidinio-3-yl phosphate

C₂₅H₅₂NO₄P (461.670).1/2H₂O

This compound is prepared in a manner analgous to Example 5 from 10.3 ml(0.11 mol) of phosphorus oxychloride, 27.0 g (0.10 mol) of octadecanol,32+40 ml of pyridine and 39.2 g (0.13 mol) of3-hydroxy-1,1-dimethylpiperidinium tosylate.

Yield: 18.7 g (40%)

Elemental analysis:

C H N calc.: 63.80% 11.35% 2.98% found: 63.38% 11.72% 2.63% 63.61%11.98% 2.61%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.35

Example 7 Hexadecyl (1,1-dimethylpiperidinio-2-yl)methyl phosphate

C₂₄H₅₀NO₄P (447.643).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 10.3ml (0.11 mol) of phosphorus oxychloride, 24.2 g (0.10 mol) ofhexadecanol, 32+40 ml of pyridine and 41.0 g (0.13 mol) of2-hydroxymethyl-1,1-dimethylpiperidinium tosylate.

Yield: 22.9 g (51%)

Elemental analysis:

C H N calc.: 63.13% 11.26% 3.07% found: 63.69% 11.73% 3.04% 63.75%11.71% 3.04%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.47

Example 8 Octadecyl (1,1-dimethylpiperidinio-2-yl)methyl phosphate

C₂₆H₅₄NO₄P (475.697).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 10.3ml (0.11 mol) of phosphorus oxychloride, 27.0 g (0.10 mol) ofoctadecanol, 32+40 ml of pyridine and 41.0 g (0.13 mol) of2-hydroxymethyl-1,1-dimethylpiperidinium tosylate.

Yield: 23.9 g (50%)

Elemental analysis:

C H N calc.: 64.43% 11.44% 2.89% found: 64.50% 11.61% 2.67% 64.11%11.49% 2.77%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.47

Example 9 Hexadecyl (1,1-dimethylpiperidinio-3-yl)methyl phosphate

C₂₄H₅₀NO₄P (447.643).1H₂O

This compound is prepared in a manner analogously to Example 5 from 10.3ml (0.11 mol) of phosphorus oxychloride, 24.2 g (0.10 mol) ofhexadecanol, 32+40 ml of pyridine and 41.0 g (0.13 mol) of3-hydroxymethyl-1,1-dimethylpiperidinium tosylate.

Yield: 17.2 g (39%)

Elemental analysis:

C H N calc.: 61.91% 11.26% 3.01% found: 62.32% 12.21% 2.86% 61.79%11.96% 2.98%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.29

Example 10 Octadecyl (1,1-dimethylpiperidinio-3-yl)methyl phosphate

C₂₆H₅₄NO₄P (475.697).H₂O

This compound is prepared in a manner analogous to Example 5 from 10.3ml (0.11 mol) of phosphorus oxychloride, 27.0 g (0.10 mol) ofoctadecanol, 32+40 ml of pyridine and 41.0 g (0.13 mol) of3-hydroxymethyl-1,1-dimethylpiperidinium tosylate.

Yield: 16.7 g (35%)

Example 12 Hexadecyl 1,1-dimethylhexahydroazepinio-4-yl phosphate

C₂₄H₄₈NO₄P (445.62)

This compound is prepared in a manner analogous to Example 5 from 10.8 g(45 mmol) of hexadecanol, 4.6 ml (50 mmol) of phosphorus oxychloride,10+20 ml of pyridine and 21.3 g (67.5 mmol) of4-hydroxy-1,1-dimethylhexahydrotosylate. It is purified by flashchromatography on silica gel with methylene chloride/methanol/25%ammonia 70:30:10.

Yield: 5.0 g (25%)

Elemental analysis:

C H N calc.: 64.69% 10.86% 3.14% found: 63.90% 11.54% 3.22% 64.08%11.59% 3.24%

Thin layer chromatogram:

(chloroform/methanol/25% ammonia 80:25:5)

Rf=0.10

(1-butanol/glacial acetic acid/water 40:10:10)

Rf=0.10

Melting point: >250° C. (decomposition)

Example 13 Octadecyl 1,1-dimethylhexahydroazepinio-4-yl phosphate

C₂₆H₅₄NO₄P (475.695).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 12.1 g(45 mmol) of octadecanol, 4.6 ml (50 mmol) of phosphorus oxychloride,10+20 ml of pyridine and 21.3 g (67.5 mmol) of4-hydroxy-1,1-dimethylhexahydroazepinium tosylate. It is purified byflash chromatography on silica gel with methylene chloride/methanol/25%ammonia 70:30:10.

Yield: 5.5 g (26%)

Elemental analysis:

C H N calc.: 64.43% 11.44% 2.89% found: 64.54% 11.64% 2.82% 64.66%11.58% 2.64%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.22

Melting point: >250° C. (decomposition)

Example 14 Cis-Δ⁹-octadecenyl 1,1-dimethylhexahydro azepinio-4-ylphosphate

C₂₆H₅₂NO₄P (473.679).H₂O

This compound is prepared in a manner analogous to Example 5 from 12.1 g(45 mmol) of cis-.9-octadecenol, 4.6 ml (50 mmol) of phosphorusoxychloride, 10+20 ml of pyridine and 21.3 g (67.5 mmol) of4-hydroxy-1,1-dimethylhexahydroazepinium tosylate. It is purified byflash chromatography on silica gel with methylene chloride/methanol/25%ammonia 70:30:10.

Yield: 4.5 g (21%)

Elemental analysis:

C H N calc.: 63.51% 11.07% 2.85% found: 64.05% 11.21% 3.10% 63.80%11.06% 3.06%

Thin layer chromatogram:

(chloroform/methanol/25% ammonia 70:40:10)

Rf=0.28

(1-butanol/glacial acetic acid/water 40:10:10)

Rf=0.10

Example 15 Eicosyl 1,1-dimethylhexahydroazepinio-4-yl phosphate

C₂₈H₅₈NO₄P (503.754).H₂O

This compound is prepared in a manner analogous to Example 5 from 13.4 g(45 mmol) of eicosanol, 4.6 ml (50 mmol) of phosphorus oxychloride,10+20 ml of pyridine and 21.3 g (67.5 mmol) of4-hydroxy-1,1-dimethylhexahydroazepinium tosylate. It is purified byflash chromatography on silica gel with methylene chloride/methanol/25%ammonia 70:30:10.

Yield: 5.7 g (25%)

Elemental analysis:

C H N calc.: 64.46% 11.59% 2.68% found: 63.51% 11.48% 2.95% 64.00%11.79% 2.91%

Thin layer chromatogram:

(chloroform/methanol/25% ammonia 70:40:10)

Rf=0.12

Example 16 Erucyl 1,1-dimethylhexahydroazepinio-4-yl phosphate

C₃₀H₆₀NO₄P (529.789).H₂O

This compound is prepared in a manner analogous to Example 5 from 16.2 g(50 mmol) of erucyl alcohol, 5.1 ml (55 mmol) of phosphorus oxychloride,18+30 ml of pyridine and 20.5 g (65 mmol) of4-hydroxy-1,1-dimethylhexahydroazepinium tosylate. It is purified byflash chromatography on silica gel with methylene chloride/methanol/25%ammonia 70:30:10.

Yield-4.1 g (15%)

Elemental analysis:

C H N calc.: 65.78% 11.41% 2.56% found: 65.76% 12.01% 2.97% 65.82%11.63% 2.96%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.30

Example 17 Octadecyl 1,1-dimethylpyrrolidinio-3-yl phosphate

C₂₄H₅₀NO₄P (447.643).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 3.25 g(12 mmol) of octadecanol, 1.21 ml (13 mmol) of phosphorus oxychloride,3.7+4.8 ml of pyridine and 4.31 g (15 mmol) ofhydroxy-1,1-dimethylpyrrolidinium tosylate. The crude product ispurified by dissolution in 96% ethanol and treatment with Amberlite MB3ion exchanger.

Yield: 1.31 g (25%)

Elemental analysis:

C H N calc.: 63.13% 11.26% 3.07% found: 62.99% 11.28% 2.80% 62.74%11.27% 2.89%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.25

Example 18 Hexadecyl 2-(1,1-dimethylpyrrolidinio-2-yl)ethyl phosphate

C₂₄H₅₀NO₄P (447.643).H₂O

This compound is prepared in a manner analogous to Example 5 from 9.21 g(38 mmol) of hexadecanol, 3.9 ml (42 mmol) of phosphorus oxychloride,13+16 ml of pyridine and 15.8 g (50 mmol) of2-(2-hydroxyethyl)-1,1-dimethylpyrrolidinium tosylate. It is purified bydissolution in 96% ethanol and treatment with Amberlite MB3 ionexchanger.

Yield: 6.0 g (35%)

Elemental analysis:

C H N calc.: 61.91% 11.26% 3.01% found: 61.82% 11.69% 3.21% 61.93%11.86% 3.28%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.38

Example 19 Octadecyl 2-(1,1-dimethylpyrrolidinio-2-yl)ethyl phosphate

C₂₆H₅₄NO₄P (475.697).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 10.3 g(38 mmol) of octadecanol, 3.9 ml (42 mmol) of phosphorus oxychloride,13+16 ml of pyridine and 15.8 g (50 mmol) of2-(2-hydroxyethyl)-1,1-dimethylpyrrolidinium tosylate. It is purified bydissolution in 96% ethanol and treatment with Amberlite MB3 ionexchanger.

Yield: 7.8 g (43%)

Elemental analysis:

C H N calc.: 64.43% 11.44% 2.89% found: 64.69% 11.77% 2.64% 64.84%11.88% 2.69%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.35

Example 20 Hexadecyl (1,1-dimethylpyrrolidinio-2-yl)methyl phosphate

C₂₃H₄₈NO₄P (433.616).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 9.21 g(38 mmol) of hexadecanol, 3.9 ml (42 mmol) of phosphorus oxychloride,13+16 ml of pyridine and 15.1 g (50 mmol) of2-hydroxymethyl-1,1-dimethylpyrrolidinium tosylate. It is purified bydissolution in 96% ethanol and treatment with Amberlite MB3 ionexchanger.

Yield: 8.3 g (51%)

Elemental analysis:

C H N calc.: 62.41% 11.16% 3.16% found: 62.09% 11.48% 3.01% 62.25%11.66% 3.09%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.33

Example 21 Octadecyl (1,1-dimethylpyrrolidinio-2-yl)methyl phosphate

C₂₅H₅₂NO₄P (461.67).1/2H₂O

This compound is prepared in a manner analogous to Example 5 from 10.3 g(38 mmol) of octadecanol, 3.9 ml (42 mmol) of phosphorus oxychloride,13+16 ml of pyridine and 15.1 g (50 mmol) of2-hydroxymethyl-1,1-dimethylpyrrolidinium tosylate. It is purified bydissolution in 96% ethanol and treatment with Amberlite MB3 ionexchanger.

Yield: 9.0 g (52%)

Elemental analysis:

C H N calc.: 63.80% 11.35% 2.98% found: 63.13% 11.57% 2.84% 63.55%11.66% 2.82%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.35

Example 22 Hexadecyl 1-methylquinuclidinio-3-yl phosphate

C₂₄H₄₈NO₄P (445.64).1.5 H₂O

2.7 ml (30 mmol) of phosphorus oxychloride are dissolved in 25 ml ofchloroform and cooled to 5-10° C. and a solution of 6.4 g (26 mmol) ofhexadecanol and 10 ml of pyridine in 50 ml of chloroform is addeddropwise over one hour. After subsequent stirring for half an hour atroom temperature, a solution of 4.5 g (35 mmol) of 3-hydroxyquinuclidineand 5 ml of pyridine in 10 ml of chloroform is added. After stirring for5 hours at room temperature, the mixture is hydrolyzed with 15 mlof-water and subsequently stirred for half an hour. It is then washedtwice with 100 ml of water/methanol (1:1) and the organic phase is driedover magnesium sulphate and concentrated to dryness. The residue ischromatographed on silica gel with methylene chloride/methanol 80:25 andthen methylene chloride/methanol/25% ammonia 80:25:5. The productfractions are purified, evaporated to dryness and crystallized withacetone. The crystals are dried under vacuum over P₂O₅.

Yield: 4.95 g (44%) of hexadecyl quinuclidinio-3-yl phosphate

4.95 g (11.5 mmol) of hexadecyl quinuclidinio-3-yl phosphate aredissolved in 30 ml of methanol, 13.7 g (69 mmol) of potassium carbonateand 8.5 ml of water are added and a solution of 3.3 ml (35 mmol) ofdimethyl sulphate in 5 ml of methanol is added dropwise, with thoroughstirring. After stirring for 14 hours at room temperature, the inorganicsalts are filtered off, the filtrate is concentrated to dryness and theresidue is taken up in methylene chloride. After filtration, thefiltrate is chromatographed on silica gel with methylenechloride/methanol/25% ammonia 70:30:5. The product fractions arecombined, evaporated to dryness and stirred with acetone untilcrystallization occurs. The crystals are dried under vacuum over P₂O₅.

Yield: 2.7 g (49%)

Elemental analysis:

C H N calc.: 60.99% 10.88% 2.96% found: 61.38% 11.04% 3.29% 61.46%11.22% 3.25%

Thin layer chromatogram:

(chloroform/methanol/25% ammonia 70:40:10)

Rf=0.44

Example 23 Octadecyl 1-methylquinuclidinio-3-yl phosphate

C₂₆H₅₂NO₄P (473.68).2H₂O

This compound is prepared in a manner analogous to Example 5 from 18.3 g(67.5 mmol) of octadecanol, 7.0 ml (75 mmol) of phosphorus oxychloride,18+20 ml of pyridine and 28.3 g (90 mmol) of3-hydroxy-1-methylquinuclidinium tosylate. It is purified by dissolutionin 96% ethanol and treatment with Amberlite MB3 ion exchanger.

Yield: 18.4 g (57%)

Elemental analysis:

C H N calc.: 61.27% 11.07% 2.75% found: 61.27% 10.91% 2.45% 61.95%11.23% 2.51%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.37

(1-butanol/glacial acetic acid/water 40:10:10)

Rf=0.13

Example 24 Hexadecyl 1,1-dimethyltropanio-4-yl phosphate

C₂₅H₅₀NO₄P (459.654).H₂O

This compound is prepared in a manner analogous to Example 5 from 12.1 g(50 mmol) of hexadecanol, 5.1 ml (55 mmol) of phosphorus oxychloride,17+40 ml of pyridine and 21.3 g (65 mmol) of4-hydroxy-1,1-dimethyltropanium tosylate. It is purified by dissolutionin 96% ethanol, treatment with Amberlite MB3 ion exchanger andrecrystallization from acetone.

Yield: 11.3 g (49%)

Elemental analysis:

C H N calc.: 62.86% 10.97% 2.93% found: 62.45% 11.52% 2.82% 62.58%11.52% 2.75%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.28

Example 25 Octadecyl 1,1-dimethyltropanio-4-yl phosphate

C₂₇H₅₄NO₄P (487.708)

This compound is prepared in a manner analogous to Example 5 from 13.5 g(50 mmol) of octadecanol, 5.1 ml (55 mmol) of phosphorus oxychloride,17+20 ml of pyridine and 21.3 g (65 mmol) of4-hydroxy-1,1-dimethyltropanium tosylate. It is purified by dissolutionin 96% ethanol and treatment with Amberlite MB3 ion exchanger.

Yield: 10.7 g (44%)

Elemental analysis:

C H N calc.: 66.49% 11.16% 2.87% found: 65.72% 11.48% 2.64% 66.27%11.78% 2.65%

Thin layer chromatogram:

(chloroform/methanol/1 M sodium acetate in 25% ammonia 70:40:10)

Rf=0.22

EXPERIMENT 1

The inventors induced mammary carcinomas in female Sprague-Dawley rats(Mollegaard Breeding Center, DK-4236 Ejby) aged 50 days by administeringa single dose of 20 mg 7,12-dimethylbenz(a)anthracene (DMBA) dissolvedin 1 ml of olive oil to each rat by gavage. The first tumor appearedapproximately one month after feeding the subject rats with DMBA.

Tumor weights were estimated on the basis of tumor volume. This wasaccomplished by palpating tumors and comparing the volumes of thepalpated tumors with the volumes of prefabricated plasticine models inthe manner taught by Druckrey, et al., “Experimentelle Beiträge zumDosis-Problem in der Krebs-Chemotherapie und zur Wirkungsweise vonEndoxan, Dtsch. Med. Wschr., 88:651 (1963). Each of the relevantplasticine models was then weighed and converted to a tumor weight bymeans of a factor reflecting the relationship between the specificweight of each tumor tissue and its corresponding plasticine model. Toensure the accuracy of this method, the inventors simultaneouslyestimated the weights of 99 individual tumors by both palpation anddirect weighing of extirpated tumors. A statistical evaluation of theresulting data indicated a correlation coefficient of 0.98.

Test rats having a total tumor weight of approximately 1 g were randomlyallocated amongst various dosage and control groups, each group having atotal of 6-7 rats. In this way, the inventors were able to ensure anapproximately equal distribution amongst the experimental groups oftumors having different latencies, total tumor weight and numbers oftumor nodes.

After separating the rats into dosage and control groups, the inventorscommenced therapy with the compounds of Examples 1, 8, 13, 20, 21 and 22of the subject application. Each compound was dissolved in 0.9% NaCl andadministered per os (stomach tube) in accordance with the regimenschedules detailed in Graphs 1-9 hereinbelow. The control group wasgiven tap water in accordance with the same schedule.

Following treatment, the test rats were observed for a period of atleast 4 days after administration of the last scheduled dose. During theobservation period, tumor weights were determined for each of the testrats at regular intervals.

The test rats were all housed under specific pathogen free (SPF)conditions with unrestricted water supply (acidified to pH 3) andstandard pellet lab chow (Altromin 1324).

EXPERIMENT 2

The inventors used female, nu/nu mice (strain NMRI) aged 9-10 weeks andweighing 21-29 g (Breeder: Bomholtgard Breeding and Research Center,DK-8680 Ry) for testing. Tumor fragments consisting of the human KBtumors (ATCC; Rockville, Md.; cell line ATCC CCL 17 KB, human epidermoidlarynx tumors), and having an average diameter of 2 mm, were implantedsubcutaneously into the right side of the test mice.

The test mice were randomly assigned to various treatment and controlgroups.

Tumor weights were estimated by first palpating the tumors and thencomparing the volumes of the palpated tumors with the volumes ofprefabricated plasticine models according to Druckrey, et al.,“Experimentelle Beiträge zum Dosis-Problem in der Krebs-Chemotherapieund zur Wirkungsweise von Endoxan, Dtsch. Med. Wschr., 88:651 (1963).After determining the weight of the models, the inventors converted eachof these values to a tumor weight by determining the relationshipbetween the specific weight of each tumor tissue and its correspondingplasticine model. To ensure the accuracy of this method, the inventorssimultaneously estimated the weights of 99 individual tumors by bothpalpation and direct weighing of extirpated tumors. A statisticalevaluation of the resulting data indicated a correlation coefficient of0.98.

Once the KB tumor implants attained a weight of approximately 0.2 g, theinventors commenced therapy with the compounds of Examples 1, 8, 17, 21,22, 24 and 25 of the subject application. Each compound was dissolved in0.9% NaCl and administered per os (stomach tube) in accordance with theregimen schedules detailed in Graphs 10-17 hereinbelow. The control micewere treated with the vehicle alone.

The mice were observed for a period of at least 21 days followingadministration of the last scheduled dose. During the observationperiod, tumor weights were determined for each of the test mice atregular intervals.

The test mice were all housed under specific pathogen free (SPF)conditions with unrestricted water supply (acidified to pH 3) andstandard pellet lab chow (Altromin 1324).

Results of Experiments 1 and 2

Results from Experiments 1 and 2 were calculated and expressed inaccordance with the growth inhibition index (GII) described in Voegeli,et al., “Selective cytostatic activity of Hexadecylphosphocholineagainst tumor cells in vitro leads to the establishment of an in vivoscreening system for phospholipid analogues,” Int. J. Oncol., 2:161(1993). The GII values from Experiments 1 and 2 are detailed in Table 1below.

TABLE 1 Example Dose DMBA KB Number (mg/kg) (G.I.I. %) (G.I.I. %) 1 1 ×511 103 4 × 100 119 14 × 31.6 108 28 × 68.1 137 8 4 × 100 125 8 2 × 316100 10 2 × 316 99 13 1 × 511 145 14 × 46.4 129 17 2 × 100 90 20 4 × 10099 21 4 × 100 116 2 × 316 106 22 4 × 100 105 2 × 383 102 24 2 × 215 10825 2 × 316 110 G.I.I. = Growth Inhibition Index (>100 = Tumorregression)

The data presented in Graphs 1-17 and Table 1 hereinabove demonstratesthat when the compounds of Examples 8, 10 and 20 are administered inaccordance with specified dosage regimens, it is possible to achieve areduction in tumor volume below that of initial tumor volume (GII=100%).It is further demonstrated by this data that when the compounds ofExamples 1, 8, 13, 21, 22, 24 and 25 of the subject application areadministered in accordance with specified dosage regimens, tumorregression is possible (GII>100%). Finally, this data shows that thecompounds of Examples 1, 21 and 25 are capable of effecting tumorremission, leading to a complete disappearance of tumors.

Treating the DMBA-induced mammary carcinomas in test rats and the KBimplanted tumors in test mice with standard cytostatics (e.g.,Cyclophosphamide, Cisplatin and Adriamycin) proved relativelyineffective. This result demonstrates that the compounds describedherein are superior to presently known cytostatics employed clinicallyin the treatment of tumors.

While the invention has been described in connection with what ispresently considered to be the most practical and preferred embodiments,it is understood that the invention is not limited to the disclosedembodiments, but on the contrary is intended to cover variousmodifications and equivalent arrangements included within the spirit andscope of the appended claims.

Thus, it is to be understood that variations in the compounds and usesthereof can be made without departing from the novel aspects of theinvention as defined in the claims.

What is claimed is:
 1. A method of treating protozoal and fungaldiseases which comprises administering to a host having a protozoal orfungal disease an effective amount of a compound of formula I

in which R is a linear or branched alkyl radical having 10 to 24 carbonatoms, which can also contain one to three double bonds or triple bonds;R₁ and R₂ independently of each other are hydrogen or a linear, branchedor cyclic hydrocarbon radical having 1 to 6 carbon atoms, thehydrocarbon radical being saturated or unsaturated and optionallycontaining a substituent selected from the group consisting of Cl, OHand NH₂, wherein two of the hydrocarbon radicals may be bonded togetherto form a ring; A is a single bond or one of the groups of the formulae:

 in which the groups having formulae (II) to (VI) are oriented in such away that the oxygen atom is bonded to the phosphorous atom of compound(I); X is an oxygen or sulfur atom or NH when A is a single bond, or anoxygen or sulfur atom when A is one of the groups having formulae (II)to (IV); y is equal to 0 or a natural number between 1 and 3; and m andn independently of one another are 0 or a natural number selected suchthat the sum of m+n equals
 4. 2. A method as set forth in claim 1 inwhich the disease is leishmaniasis.
 3. A method of treating psoriasiscomprising administering to a patient having psoriasis an effectiveamount of a compound of formula I

in which R is a linear or branched alkyl radical having 10 to 24 carbonatoms, which can also contain one to three double bonds or triple bonds;R₁ and R₂ independently of each other are hydrogen or a linear, branchedor cyclic hydrocarbon radical having 1 to 6 carbon atoms, thehydrocarbon radical being saturated or unsaturated and optionallycontaining a substituent selected from the group consisting of Cl, OHand NH₂, wherein two of the hydrocarbon radicals may be bonded togetherto form a ring; A is a single bond or one of the groups of the formulae:

 in which the groups having formulae (II) to (VI) are oriented in such away that the oxygen atom is bonded to the phosphorous atom of compound(I); X is an oxygen or sulfur atom or NH when A is a single bond, or anoxygen or sulfur atom when A is one of the groups having formulae (II)to (IV); y is equal to 0 or a natural number between 1 and 3; and m andn independently of one another are 0 or a natural number selected suchthat the sum of m+n equals
 4. 4. A method of treating bone marrow damagedue to treatment with cytostatic agents and other myelotoxic activeingredients which comprises administering, to a host having bone marrowdamage due to treatment with cytostatic agents or other myelotoxicactive ingredients, an effective amount of a compound of formula I

in which R is a linear or branched alkyl radical having 10 to 24 carbonatoms, which can also contain one to three double bonds or triple bonds;R₁ and R₂ independently of each other are hydrogen or a linear, branchedor cyclic hydrocarbon radical having 1 to 6 carbon atoms, thehydrocarbon radical being saturated or unsaturated and optionallycontaining a substituent selected from the group consisting of Cl, OHand NH₂, wherein two of the hydrocarbon radicals may be bonded togetherto form a ring; A is a single bond or one of the groups of the formulae:

 in which the groups having formulae (II) to (VI) are oriented in such away that the oxygen atom is bonded to the phosphorous atom of compound(I); X is an oxygen or sulfur atom or NH when A is a single bond, or anoxygen or sulfur atom when A is one of the groups having formulae (II)to (IV); y is equal to 0 or a natural number between 1 and 3; and m andn independently of one another are 0 or a natural number selected suchthat the sum of m+n equals 4.